A study of transcriptome in leaf rust infected bread wheat involving seedling resistance gene Lr28

Leaf rust disease causes severe yield losses in wheat throughout the world. During the present study, high-throughput RNA-Seq analysis was used to gain insights into the role of Lr28 gene in imparting seedling leaf rust resistance in wheat. Differential expression analysis was conducted using a pair of near-isogenic lines (NILs) (HD 2329 and HD 2329 + Lr28) at early (0 h before inoculation (hbi), 24 and 48 h after inoculation (hai)) and late stages (72, 96 and 168 hai) after inoculation with a virulent pathotype of pathogen Puccinia triticina. Expression of a large number of genes was found to be affected due to the presence/absence of Lr28. Gene ontology analysis of the differentially expressed transcripts suggested enrichment of transcripts involved in carbohydrate and amino acid metabolism, oxidative stress and hormone metabolism, in resistant and/or susceptible NILs. Genes encoding receptor like kinases (RLKs) (including ATP binding; serine threonine kinases) and other kinases were the most abundant class of genes, whose expression was affected. Genes involved in reactive oxygen species (ROS) homeostasis and several genes encoding transcription factors (TFs) (most abundant being WRKY TFs) were also identified along with some ncRNAs and histone variants. Quantitative real-time PCR was also used for validation of 39 representative selected genes. In the long term, the present study should prove useful in developing leaf rust resistant wheat cultivars through molecular breeding.

Lr72 Confers Resistance to Leaf Rust in Durum Wheat Cultivar Atil C2000

Leaf rust, caused by Puccinia triticina (Pt), has become a globally important disease for durum wheat (Triticum turgidum subsp. durum) since the detection of race group BBG/BN, which renders ineffective a widely deployed seedling resistance gene present in several popular cultivars including Mexican cultivars Altar C84 and Atil C2000. The resistance gene continues to play a key role in protecting durum wheat against bread wheat–predominant races since virulence among this race group has not been found. We developed F3 and F5 mapping populations from a cross between Atil C2000 and the susceptible line Atred #1. Resistance was characterized by greenhouse seedling tests using three Pt races. Segregation tests indicated the presence of a single gene, which was mapped to the distal end of 7BS by bulk segregant analysis. The closest marker, wmc606, was located 5.5 cM proximal to the gene. No known leaf rust resistance genes are reported in this region; this gene was therefore designated as Lr72. The presence of Lr72 was further investigated in greenhouse tests in a collection of durum wheat using 13 Pt races. It was concluded that at least one additional gene protects durum wheat from bread wheat–predominant Pt races.

Analysis of differentially expressed genes in leaf rust infected bread wheat involving seedling resistance gene Lr28

Genome-wide transcriptome analysis of seedling resistance to leaf rust conferred by Lr28 gene in wheat (Triticum aestivum L.) was conducted to identify differentially expressed genes during incompatible interaction. A virulent leaf rust race 77–5 was used for inoculation of resistant (HD2329 + Lr28) and susceptible (HD2329 – Lr28) wheat NILs and cDNA-AFLP analyses was carried out. As many as 223 differential transcripts appeared following leaf rust inoculation; these included 122 transcripts that appeared exclusively in resistant NIL, whereas 39 transcripts appeared both in resistant and susceptible NILs. Sequence analyses of 37 transcripts, which appeared in the resistant NIL revealed that 15 transcripts had homology with genes involved in protein synthesis, signal transduction, transport, disease resistance and metabolism. The functions of remaining 22 transcripts could not be determined; these included six novel genes reported for the first time in wheat. Specific primers could be designed for 18 of the 37 transcripts, which included genes with putative and unknown functions. Quantitative real time PCR analysis was conducted using these 18 pairs of primers. A majority (13) of these transcripts appeared within 48 h reaching a peak value at 96 h in resistant NIL signifying their role in providing leaf rust resistance.